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食品与发酵工业  2019, Vol. 45 Issue (1): 94-99    DOI: 10.13995/j.cnki.11-1802/ts.016282
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
脂质氢过氧化物氧化对核桃分离蛋白结构的影响
王丹丹,孙领鸽,毛晓英*
(石河子大学食品学院,新疆 石河子,832000)
Effects of lipid oxidative modification by hydroperoxides on the structure of walnut protein
WANG Dandan, SUN Linge, MAO Xiaoying*
(Food College of Shihezi University, Shihezi 832000,China)
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摘要 采用脂肪氧合酶催化亚油酸构建脂质氢过氧化物——核桃分离蛋白氧化体系,研究脂质氧化反应中产生的氢过氧化物对核桃分离蛋白结构特性的影响。结果显示,向100 mL 0.05 mg/mL蛋白溶液中添加9 mL亚油酸,核桃分离蛋白溶解度下降至6.89%,游离巯基含量由2.82 μmol SH/g下降至1.92 μmol SH/g,羰基含量由2.26 nmol/mg增加至4.23 nmol/mg,表明氧化后核桃蛋白理化特性产生变化。圆二色谱分析表明,蛋白质氧化导致核桃分离蛋白α-螺旋和β-折叠含量下降,β-转角和无规则卷曲含量上升。随着亚油酸添加量的增加,蛋白质氧化程度加大,核桃分离蛋白表面疏水性从429.66下降到 405.24。内源荧光最大荧光峰位红移,内源荧光强度下降,此结果表明,脂肪氧合酶(lipoxygenase, LOX)催化亚油酸氧化诱导核桃分离蛋白聚集,并进一步使蛋白质二级和三级结构发生变化。体积凝胶色谱法结果显示,核桃分离蛋白氧化后,小分子多肽含量下降,高分子量聚集体含量增加,说明脂质氢过氧化物氧化修饰导致核桃蛋白结构改变,形成氧化聚集体。
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王丹丹
孙领鸽
毛晓英
关键词:  脂质氢过氧化物  核桃蛋白  氧化  结构    
Abstract: Walnut protein isolates were oxidized by lipid hydroperoxides, which were produced from different levels of linoleic acid oxidation catalysed by lipoxygenase. Effects of hydroperoxides on structural characteristics of walnut protein isolates were studied. The results showed that when 9 mL linoleic acid was added to 100 mL 0.05 mg/mL protein solution, walnut protein solubility decreased to 6.89%. The level of free sulfhydryl group decreased from 2.82 umol SH/g to 1.92 umol SH/g, and the carbonyl level increased from 2.26 nmol/mg to 4.23 nmol/mg. This indicated that the physicochemical properties of isolated walnut protein changed after oxidation. The CD spectrum analysis showed that protein oxidation caused reduction in the ɑ-helix and β-sheet contents, and the contents of β-turn and random-coil increased. With increasing amount of linoleic acid added, the degree of protein oxidation increased and protein surface hydrophobicity decreased from 429.66 to 405.24. Moreover, red shift happened to the highest fluorescent peak, and the intensity of endogenous fluorescence decreased. This suggested that lipid hydroperoxide oxidation could induce aggregation of isolated walnut protein, thus leading to further changes in its secondary and tertiary structures. Furthermore, oxidation caused the contents of small molecule polypeptides decreasing, and the levels of protein aggregates gradually increased in the molecular weight distribution of walnut protein. The results indicated that the structure of walnut protein changed by lipid hydroperoxide oxidation, which caused the formation of oxidative aggregates of walnut protein.
Key words:  lipid hydroperoxide    walnut protein    oxidation    structure
收稿日期:  2017-11-15                出版日期:  2019-01-15      发布日期:  2019-02-01      期的出版日期:  2019-01-15
基金资助: 国家自然科学基金(31560433)
作者简介:  硕士研究生(毛晓英副教授为通讯作者,E-mail:maoxiaoying99@163.com)。
引用本文:    
王丹丹,孙领鸽,毛晓英. 脂质氢过氧化物氧化对核桃分离蛋白结构的影响[J]. 食品与发酵工业, 2019, 45(1): 94-99.
WANG Dandan,SUN Linge,MAO Xiaoying. Effects of lipid oxidative modification by hydroperoxides on the structure of walnut protein[J]. Food and Fermentation Industries, 2019, 45(1): 94-99.
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http://sf1970.cnif.cn/CN/10.13995/j.cnki.11-1802/ts.016282  或          http://sf1970.cnif.cn/CN/Y2019/V45/I1/94
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